title: Molecular cloning and functional characterization of Argonaute-3 gene from Penaeus monodon.
abstract: Argonaute (Ago) proteins play a crucial role in the shrimp RNA interference pathway. In this study, we identified and characterized a novel Ago gene from black tiger shrimp, Penaeus monodon. The complete open reading frame of P. monodon Ago3 (PmAgo3) consisted of 2559 nucleotides encoding a polypeptide of 852 amino acids with a predicted molecular weight of 97 kDa and an isoelectric point of 9.42. Analysis of the deduced amino acid sequence of PmAgo3 revealed the presence of two signature domains of the proteins in Argonaute family including PAZ and PIWI. Phylogenetic analysis indicated that PmAgo3 is classified into Ago subfamily and shared the highest amino acid sequence identity (83%) with Litopenaeus vannamei Ago2. Monitoring of the PmAgo3 expression by quantitative real-time PCR revealed that this gene was significantly up-regulated following dsRNA administration, while no significant difference in its expression was observed following yellow head virus (YHV) challenge. In contrast, inhibition of YHV mRNA expression was observed in PmAgo3-knockdown shrimp. These data imply that PmAgo3 is involved in the dsRNA-mediated gene silencing mechanism and plays an important role in YHV replication in the black tiger shrimp.
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Phutthamonthon 4 Road, Salaya, Nakhon Pathom 73170, Thailand.
authors.forename: Amnat
authors.initials: A
authors.lastname: Phetrungnapha
authors.forename: Teerapong
authors.initials: T
authors.lastname: Ho
authors.forename: Apinunt
authors.initials: A
authors.lastname: Udomkit
authors.forename: Sakol
authors.initials: S
authors.lastname: Panyim
authors.forename: Chalermporn
authors.initials: C
authors.lastname: Ongvarrasopone
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D005786:Gene Expression Regulation / Q000502:physiology*; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity
chemical_list: D060565:Argonaute Proteins; D012333:RNA, Messenger
keywords: Argonaute; Black tiger shrimp; Double-stranded RNA; RNAi; YHV